Fiber a2000i Moa
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Introducing the Automated Fiber Analyzer• Performs Crude Fiber, ADF and NDF determinations
• Automatically adds required solutions and hot water rinses
• Batch Process up to 24 samples at one time
• Accurate and Precise results, time after time
• Computerized controls eliminate technician variability
The ANKOM2000 Fiber Analyzer automates fiber determinations.Capable of Acid Detergent, Neutral Detergent and Crude Fiber analysis,the ANKOM2000 eliminates chemical and hot water handling. The samples are prepared using ANKOM Filter Bag Technology and placed ina bag suspender. At that point the technician simply selects the analytical method on the computer controller and pushes a button. Thesystem automatically adds the appropriate chemicals and performs thenecessary rinses. The solution is heated and solublization takes placewithout manual intervention. When the process is complete, the technician simply removes the bags and completes a final drying beforeweighing the samples.
Helping To Feed The World!
ANKOMTechnology isthe developer ofFilter Bag
Technology (FBT).Samples are complete-
ly encapsulated which insures easeof handling and virtually elimi-nates error. Operator exposure tochemicals is eliminated. UsingFilter Bags, productivity soars assamples are batch processed in anautomated system. The samplesper day your lab can process willdramatically increase. Costs aredecreased as your technicians arefree to take on other tasks whilethe ANKOM automated analyzerhandles the process.
• Accurate Results• Increased Precision• Batch Processing• Lower Costs• High Output
Used in over 85 countries around the world, theANKOM200 Fiber Analyzer provides a low cost alternative for the laboratory seeking increased efficiency. Technicians simply add the required solutions and hot water rinses according to the method and the ANKOM200 handles the rest.
Also Available
ANKOM_SELL_SHEETS_A2000_V14 6/19/07 3:59 PM Page 1
SpecificationsSample SpecificationsSample Size 0.5 – 1.0 gramFiber Range: 0% - 100%Samples per batch: up to 24
Samples per day Acid Detergent Fiber = 144Neutral Detergent Fiber = 120 Crude Fiber = 96
Instrument Specifications: ANKOM2000 ANKOM200
Dimensions: 18”w x 14.5”d x 23”h 16”w x 9”d x 20”hOperating Temperature 100°C 100°CPower Requirements 120 – 240V, 50/60 Hz 120–240V, 50/60 HzWeight 46 kg. 20 kg
ANKOM Technology2052 O'Neil Rd.Macedon, NY 14502
Phone: (315) 986 - 8090 Fax: (315) 986 - 8091 E-mail: [email protected]
ANKOM Technology is the developer of Filter Bag Technology (FBT) used aroundthe world for fiber and fat analysis. With customers in over 85 countries, ANKOMhas a reputation for quality and innovation. Constantly seeking to develop bettermethods for time consuming analytical methods, ANKOM Technology focuses oncustomers needs. We offer instruments, chemicals and other ancillary products tosupport fiber studies, crude fat extraction, total fat extraction, in vitro and in situresearch and more. We work hard to keep costs low with quality and service high.
Helping To Feed The World!
ANKOM_SELL_SHEETS_A2000_V14 6/19/07 4:00 PM Page 2
Introducing the Automated Fiber Analyzer• Performs Crude Fiber, ADF and NDF determinations
• Automatically adds required solutions and hot water rinses
• Batch Process up to 24 samples at one time
• Accurate and Precise results, time after time
• Computerized controls eliminate technician variability
The ANKOM2000 Fiber Analyzer automates fiber determinations.Capable of Acid Detergent, Neutral Detergent and Crude Fiber analysis,the ANKOM2000 eliminates chemical and hot water handling. The samples are prepared using ANKOM Filter Bag Technology and placed ina bag suspender. At that point the technician simply selects the analytical method on the computer controller and pushes a button. Thesystem automatically adds the appropriate chemicals and performs thenecessary rinses. The solution is heated and solublization takes placewithout manual intervention. When the process is complete, the technician simply removes the bags and completes a final drying beforeweighing the samples.
Helping To Feed The World!
ANKOMTechnology isthe developer ofFilter Bag
Technology (FBT).Samples are complete-
ly encapsulated which insures easeof handling and virtually elimi-nates error. Operator exposure tochemicals is eliminated. UsingFilter Bags, productivity soars assamples are batch processed in anautomated system. The samplesper day your lab can process willdramatically increase. Costs aredecreased as your technicians arefree to take on other tasks whilethe ANKOM automated analyzerhandles the process.
• Accurate Results• Increased Precision• Batch Processing• Lower Costs• High Output
Used in over 85 countries around the world, theANKOM200 Fiber Analyzer provides a low cost alternative for the laboratory seeking increased efficiency. Technicians simply add the required solutions and hot water rinses according to the method and the ANKOM200 handles the rest.
Also Available
ANKOM_SELL_SHEETS_A2000_V14 6/19/07 3:59 PM Page 1
SpecificationsSample SpecificationsSample Size 0.5 – 1.0 gramFiber Range: 0% - 100%Samples per batch: up to 24
Samples per day Acid Detergent Fiber = 144Neutral Detergent Fiber = 120 Crude Fiber = 96
Instrument Specifications: ANKOM2000 ANKOM200
Dimensions: 18”w x 14.5”d x 23”h 16”w x 9”d x 20”hOperating Temperature 100°C 100°CPower Requirements 120 – 240V, 50/60 Hz 120–240V, 50/60 HzWeight 46 kg. 20 kg
ANKOM Technology2052 O'Neil Rd.Macedon, NY 14502
Phone: (315) 986 - 8090 Fax: (315) 986 - 8091 E-mail: [email protected]
ANKOM Technology is the developer of Filter Bag Technology (FBT) used aroundthe world for fiber and fat analysis. With customers in over 85 countries, ANKOMhas a reputation for quality and innovation. Constantly seeking to develop bettermethods for time consuming analytical methods, ANKOM Technology focuses oncustomers needs. We offer instruments, chemicals and other ancillary products tosupport fiber studies, crude fat extraction, total fat extraction, in vitro and in situresearch and more. We work hard to keep costs low with quality and service high.
Helping To Feed The World!
ANKOM_SELL_SHEETS_A2000_V14 6/19/07 4:00 PM Page 2
ADF Method
Method 12
11-12-13 pg. 1 of 2
Acid Detergent Fiber in Feeds - Filter Bag Technique (for A2000 and A2000I)
Definition
This method determines Acid Detergent Fiber, which is the residue remaining after digesting with H2SO4 and CTAB. The fiber residues are
predominantly cellulose and lignin.
Scope
This method is applicable to grains, feeds, forages, and all fiber-bearing material.
Apparatus
1. Analytical Balance—capable of weighing 0.1 mg.
2. Oven—capable of maintaining a temperature of 102 ± 2°C.
3. Digestion instrument—capable of performing the digestion at
100 ± 0.5°C and maintaining a pressure of 10-25psi. The
instrument must be capable of creating a similar flow around
each sample to ensure uniformity of extraction (ANKOM2000
with 65rpm agitation, ANKOM Technology).
4. Filter Bags—constructed from chemically inert and heat resistant
filter media, capable of being heat sealed closed and able to
retain 25 micron particles while permitting solution penetration
(F57 and F58, ANKOM Technology).
5. Heat sealer—sufficient for sealing the filter bags closed to ensure
complete closure (1915, ANKOM Technology).
6. Desiccant Pouch—collapsible sealable pouch with desiccant
inside that enables the removal of air from around the filter bags
(MoistureStop weigh pouch, ANKOM Technology).
7. Marking pen—solvent and acid resistant (F08, ANKOM
Technology).
Reagents
1. Acid Detergent Solution—Add 20g cetyl trimethylammonium
bromide (CTAB) to 1L 1.00N H2SO4 previously standardized
(premixed chemical solution available from ANKOM). Agitate
and heat to aid solution.
CAUTION1: Sulfuric acid is a strong acid and will cause severe
burns. Protective clothing should be worn when working with
this acid. Always add acid to water and not the reverse.
CAUTION2: CTAB will irritate mucous membranes. A dust
mask and gloves should be worn when handling this chemical.
Sample Preparation
Grind samples in a centrifugal mill with a 2mm screen or cutter type
(Wiley) mill with a 1mm screen. Samples ground finer may have
particle loss from the filter bags and result in low values.
ADF Procedure (see the ADF Analysis section of the Operator’s Manual for
more detail)
1. Use a solvent resistant marker to label the filter bags to be used in
the analysis.
2. Weigh and record the weight of each empty filter bag (W1) and
zero the balance. NOTE: Do not pre-dry filter bags. Any moisture
will be accounted for by the blank bag correction.
3. Place 0.45 – 0.50g of prepared sample in up to 23 of the bags and
record the weight (W2) of each. Avoid placing the sample in the
upper 4mm of the bag.
4. Include at least one empty bag in the run to determine the
blank bag correction (C1).
NOTE: A running average blank bag correction factor (C1)
should be used in the calculation of fiber. The inclusion of at
least one blank bag in each run is mainly used as an indicator
of particle loss. A C1 larger than 1.0000 indicates that sample
particles were lost from filter bags and deposited on the blank
bag during the extraction. Any fiber particle loss from the
filter bags will generate erroneous results. If particle loss is
observed then the grinding method needs to be evaluated.
5. Using a heat sealer, completely seal each filter bag closed
within 4mm of the top to encapsulate the sample. NOTE: Use
sufficient heat to completely seal the filter bags and allow
enough cool time (2 sec) before removing each bag from the
heat sealer.
6. Pre-extract only samples containing >5% fat: Extract
samples by placing bags with samples into a container with a
top. Pour enough acetone into the container to cover the bags
and secure the top.
CAUTION3: Acetone is extremely flammable. Avoid static
electricity and use a fume hood when handling.
Shake the container 10 times and allow bags to soak for 10
minutes. Repeat with fresh acetone. Pour out acetone and
place bags on a wire screen to air-dry.
Exception – Roasted soybean: Due to the processing of
roasted soy a modification to the extraction is required. Place
roasted soy samples into a container with a top. Pour enough
acetone into the container to cover the bags and secure the
top. Shake the container 10 times and pour off the acetone.
Add fresh acetone and allow samples to soak for twelve
hours. After the soak time, pour out the acetone and place the
bags on a wire screen to air-dry.
7. Spread the sample uniformly inside the filter bags by shaking
and flicking the bags to eliminate clumping.
8. Place up to 3 bags on each of eight Bag Suspender Trays
(maximum of 24 bags). Stack the trays on the center post of
the Bag Suspender with each level rotated 120 degrees in
relation to the tray below it. Place the empty 9th tray on top.
NOTE: All nine trays must be used regardless of the number
of bags being processed.
9. Verify that the hot water supply is on and the drain hose is
securely positioned in the drain.
10. Read the Temperature Controller on the right side of the
instrument. If the temperature is higher than 20°C, cool the
Vessel as follows:
a. Fill the Vessel with cold water.
b. When the Temperature Controller reads 20°C, run
the Flush Procedure to drain the water.
c. Repeat steps a and b if necessary.
(Procedure continued on next page.)
ADF Method
Method 12
11-12-13 pg. 2 of 2
Calculations
% ADF (as-received basis) = 100 x (W3 – (W1 x C1))
W2
Where: W1 = Bag tare weight W2 = Sample weight
W3 = Dried weight of bag with fiber after extraction process
C1 = Blank bag correction (running average of final oven-dried weight divided by original blank bag weight)
ADF Procedure (continued)
11. If you are using Cubetainers for your chemicals, attach the
AD solution hose to the Cubetainer and then to Port B on the
instrument.
12. Open the Vessel Lid and insert the Bag Suspender with bags
into the Vessel and place the Bag Suspender Weight on top of
the empty 9th tray to keep the Bag Suspender submerged.
13. Follow the instructions on the ANKOM2000 display:
a. Select ADF.
b. Close the Vessel Lid.
c. Confirm hot water is on (>70°C).
d. Press START.
14. When the ADF extraction and rinsing procedures are
complete, open the Vessel Lid and remove the filter bags.
Gently press out excess water from the bags. Place bags in a
250ml beaker and add enough acetone to cover bags and soak
for 3-5 minutes.
15. Remove the filter bags from the acetone and place them on a
wire screen to air-dry. Completely dry in an oven at 102 ±
2°C. (In most ovens the filter bags will be completely dry
within 2-4 hours.) NOTE: Do not place bags in the oven until
the acetone has completely evaporated.
16. Remove the filter bags from the oven and immediately place
them directly into a collapsible desiccant pouch and flatten to
remove any air. Cool to ambient temperature and weigh the
filter bags (W3). NOTE: Do not use a conventional desiccator
container.
NDF Method
Method 13
11/13/13 pg. 1 of 2
Neutral Detergent Fiber in Feeds - Filter Bag Technique (for A2000 and A2000I)
Definition
This method determines Neutral Detergent Fiber, which is the residue remaining after digesting in a detergent solution. The fiber residues are
predominantly hemicellulose, cellulose, and lignin.
Scope
This method is applicable to grains, feeds, forages, and all fiber-bearing material.
Apparatus
1. Analytical Balance—capable of weighing 0.1 mg.
2. Oven—capable of maintaining a temperature of 102 ± 2°C.
3. Digestion instrument—capable of performing the digestion at
100 ± 0.5°C and maintaining a pressure of 10-25psi. The
instrument must be capable of creating a similar flow around
each sample to ensure uniformity of extraction (ANKOM2000
with 65rpm agitation, ANKOM Technology).
4. Filter Bags—constructed from chemically inert and heat resistant
filter media, capable of being heat sealed closed and able to
retain 25 micron particles while permitting solution penetration
(F57 and F58, ANKOM Technology).
5. Heat sealer—sufficient for sealing the filter bags closed to ensure
complete closure (1915, ANKOM Technology).
6. Desiccant Pouch—collapsible sealable pouch with desiccant
inside that enables the removal of air from around the filter bags
(MoistureStop weigh pouch, ANKOM Technology).
7. Marking pen—solvent and acid resistant (F08, ANKOM
Technology).
Reagents
1. Neutral Detergent Solution—Add 30g Sodium dodecyl sulfate
(USP), 18.61g Ethylenediaminetetraacetic disodium salt
(dehydrate), 6.81g Sodium borate, 4.56g Sodium phosphate
dibasic (anhydrous), and 10.0ml Triethylene glycol to 1L
distilled H2O (premixed chemical solution available from
ANKOM Technology). Check that pH is from 6.9 to 7.1. Agitate
and heat to aid solution.
CAUTION1: Powdered chemicals will irritate mucous
membranes. A dust mask and gloves should be worn when
handling these chemicals.
2. Alpha-amylase—Heat-stable bacterial alpha-amylase: activity =
17,400 Liquefon Units / ml (FAA, ANKOM Technology).
3. Sodium sulfite—Na2SO3, anhydrous (FSS, ANKOM
Technology)
Sample Preparation
Grind samples in a centrifugal mill with a 2mm screen or cutter type
(Wiley) mill with a 1mm screen. Samples ground finer may have
particle loss from the filter bags and result in low values.
NDF Procedure (see the NDF Analysis section of the Operator’s Manual for
more detail)
1. Use a solvent resistant marker to label the filter bags to be used in
the analysis.
2. Weigh and record the weight of each empty filter bag (W1) and
zero the balance. NOTE: Do not pre-dry filter bags. Any moisture
will be accounted for by the blank bag correction.
3. Place 0.45 – 0.50g of prepared sample in up to 23 of the bags and
record the weight (W2) of each. Avoid placing the sample in the
upper 4mm of the bag.
4. Include at least one empty bag in the run to determine the
blank bag correction (C1).
NOTE: A running average blank bag correction factor (C1)
should be used in the calculation of fiber. The inclusion of at
least one blank bag in each run is mainly used as an indicator
of particle loss. A C1 larger than 1.0000 indicates that sample
particles were lost from filter bags and deposited on the blank
bag during the extraction. Any fiber particle loss from the
filter bags will generate erroneous results. If particle loss is
observed then the grinding method needs to be evaluated.
5. Using a heat sealer, completely seal each filter bag closed
within 4mm of the top to encapsulate the sample. NOTE: Use
sufficient heat to completely seal the filter bags and allow
enough cool time (2 sec) before removing each bag from the
heat sealer.
6. Pre-extract only samples containing >5% fat: Extract
samples by placing bags with samples into a container with a
top. Pour enough acetone into the container to cover the bags
and secure the top.
CAUTION2: Acetone is extremely flammable. Avoid static
electricity and use a fume hood when handling.
Shake the container 10 times and allow bags to soak for 10
minutes. Repeat with fresh acetone. Pour out acetone and
place bags on a wire screen to air-dry.
Exception – Roasted soybean: Due to the processing of
roasted soy a modification to the extraction is required. Place
roasted soy samples into a container with a top. Pour enough
acetone into the container to cover the bags and secure the
top. Shake the container 10 times and pour off the acetone.
Add fresh acetone and allow samples to soak for twelve
hours. After the soak time, pour out the acetone and place the
bags on a wire screen to dry.
7. Spread the sample uniformly inside the filter bags by shaking
and flicking the bags to eliminate clumping.
8. Place up to 3 bags on each of eight Bag Suspender Trays
(maximum of 24 bags). Stack the trays on the center post of
the Bag Suspender with each level rotated 120 degrees in
relation to the tray below it. Place the empty 9th tray on top.
NOTE: All nine trays must be used regardless of the number
of bags being processed.
9. Verify that the hot water supply is on and the drain hose is
securely positioned in the drain.
10. If you are using Cubetainers for your chemicals, attach the
ND solution hose to the Cubetainer and then to Port A on the
instrument.
11. Open the Vessel Lid and insert the Bag Suspender with bags
into the Vessel and place the Bag Suspender weight on top of
the empty 9th tray to keep the Bag Suspender submerged.
(Procedure continued on next page.)
NDF Method
Method 13
11/13/13 pg. 2 of 2
Calculations
% NDF (as-received basis) = 100 x (W3 – (W1 x C1))
W2
Where: W1 = Bag tare weight W2 = Sample weight
W3 = Dried weight of bag with fiber after extraction process
C1 = Blank bag correction (running average of final oven-dried weight divided by original blank bag weight)
NDF Procedure (continued)
12. Follow the instructions on the ANKOM2000 display:
a. Select NDF. (Wait to close the Vessel Lid.)
b. Confirm hot water is on (>70°C).
c. Press START.
d. After the ND solution has been automatically
inserted and agitation begins, manually add 20g of
Na2SO3 and 4.0ml of alpha-amylase.
e. Close the Vessel Lid.
13. Attach the Amylase Dispenser Assembly to Port B on the
instrument. Add 8.0ml of alpha-amylase and enough water to
fill the dispenser. The ANKOM2000 will automatically add the
amylase solution to the first and second rinse.
14. When the NDF extraction and rinsing procedures are
complete, open the Vessel Lid and remove the filter bags.
Gently press out excess water from the bags. Place bags in a
250ml beaker and add enough acetone to cover bags and soak
for 3-5 minutes.
15. Remove the filter bags from the acetone and place them on a
wire screen to air-dry. Completely dry in an oven at 102 ±
2°C. (In most ovens the filter bags will be completely dry
within 2-4 hours.) NOTE: Do not place bags in the oven until
the acetone has completely evaporated.
16. Remove the filter bags from the oven and immediately place
them directly into a collapsible desiccant pouch and flatten to
remove any air. Cool to ambient temperature and weigh the
filter bags (W3). NOTE: Do not use a conventional countertop
or cabinet desiccator.
Crude Fiber Method
Method 1
11/26/13 pg. 1 of 3
Crude Fiber Analysis in Feeds - Filter Bag Technique (for A2000 and A2000I) AOAC Approved Procedure Ba 6a-05
Definition
This method determines Crude Fiber which is the organic residue remaining after digesting with 0.255N H2SO4 and 0.313N NaOH. The
compounds removed are predominantly protein, sugar, starch, lipids and portions of both the structural carbohydrates and lignin.
Scope
This method is applicable for all feed materials such as grains, meals, pet foods, mixed feeds, forages, and the following oilseeds: corn and
soybeans.
Apparatus
1. Analytical Balance—capable of weighing 0.1 mg.
2. Oven—capable of maintaining a temperature of 102 ± 2°C.
3. Electric muffle furnace—with rheostat control and pyrometer
that will maintain a temperature of 600 ± 15°C.
4. Digestion instrument—capable of performing the digestion at
100 ± 0.5°C and maintaining a pressure of 10-25psi. The
instrument must be capable of creating a similar flow around
each sample to ensure uniformity of extraction (ANKOM2000
with 65rpm agitation, ANKOM Technology).
5. Filter Bags—constructed from chemically inert and heat resistant
filter media, capable of being heat sealed closed and able to
retain 25 micron particles while permitting solution penetration
(F57 or F58, ANKOM Technology). See Numbered Notes 1.
6. Heat sealer—sufficient for sealing the filter bags closed to ensure
complete closure (1915, ANKOM Technology).
7. Desiccant Pouch—collapsible sealable pouch with desiccant
inside that enables the removal of air from around the filter bags
(MoistureStop weigh pouch, ANKOM Technology).
8. Marking pen—solvent and acid resistant (F08, ANKOM
Technology).
Reagents
1. Sulfuric acid solution—0.255 ± 0.005N. 1.25g H2SO4/100ml.
Concentration must be checked by titration.
CAUTION1: Sulfuric acid is a strong acid and will cause severe
burns. Protective clothing should be worn when working with
this acid. Always add acid to water and not the reverse.
2. Sodium hydroxide solution—0.3130 ± 005N. 1.25g
NaOH/100ml. Concentration must be checked by titration.
CAUTION2: Sodium hydroxide can severely burn the skin,
eyes, and respiratory tract. Protective clothing should be worn
when working with this acid. Always add caustic material to
water and not the reverse.
Sample Preparation
Grind samples in a centrifugal mill with a 2mm screen or cutter type
(Wiley) mill with a 1mm screen. Samples ground finer (fiber particles
less than 25 microns) may have particle loss through the filter bags
that result in lower fiber values (up to 0.5% units).
Precision
Results of the collaborative study (see Tables 1&2) indicate the
precision (Sr, RSDr, r) that the analyst should use as a benchmark for
evaluating replication in the same laboratory.
Crude Fiber Procedure (see the Crude Fiber Analysis section of the
Operator’s Manual for more detail)
1. Use a solvent resistant marker to label the filter bags to be
used in the analysis.
2. Weigh and record the weight of each empty filter bag (W1)
and zero the balance. NOTE: Do not pre-dry filter bags. Any
moisture will be accounted for by the blank bag correction.
3. Place 0.95 – 1.00g of prepared sample in up to 23 of the
bags and record the weight (W2) of each. Avoid placing the
sample in the upper 4mm of the bag.
4. Include at least one empty bag in the run to determine the
blank bag correction (C1).
NOTE: A running average blank bag correction factor (C1)
should be used in the calculation of fiber. The inclusion of at
least one blank bag in each run is mainly used as an
indicator of particle loss. A C1 larger than 1.0000 indicates
that sample particles were lost from filter bags and deposited
on the blank bag. Any fiber particle loss from the filter bags
will generate erroneous results. If particle loss is observed
then the grinding method needs to be evaluated.
5. Using a heat sealer, completely seal each filter bag closed
within 4mm of the top to encapsulate the sample. NOTE:
Use sufficient heat to completely seal the filter bags and
allow enough cool time (2 sec) before removing each bag
from the heat sealer.
6. Extract fat from samples by placing all bags into a 250ml
container. Add enough petroleum ether to cover bags and
soak for 10 minutes.
CAUTION3: Petroleum ether is extremely flammable.
Avoid static electricity. A fume hood should be used at all
times when using petroleum ether.
Pour off the solvent and allow the bags to air-dry. Spread the
sample uniformly inside the filter bags by shaking and
flicking the bags to eliminate clumping.
7. Place up to 3 bags on each of eight Bag Suspender Trays
(maximum of 24 bags). Stack the trays on the center post of
the Bag Suspender with each level rotated 120 degrees in
relation to the tray below it. Place the empty 9th tray on top.
NOTE: All nine trays must be used regardless of the number
of bags being processed.
8. Verify that the hot water supply is on and the drain hose is
securely positioned in the drain.
(Procedure continued on next page.)
Crude Fiber Method
Method 1
11/26/13 pg. 2 of 3
Calculations
% Crude Fiber = 100 x (W3 – (W1 x C1))
W2
Where: W1 = Bag tare weight W2 = Sample weight
W3 = Weight of Organic Matter (loss of weight on ignition of bag and fiber)
C1 = Ash corrected blank bag factor (running average of loss of weight on ignition of blank bag/original blank bag)
Crude Fiber Procedure (continued)
9. Read the Temperature Controller on the right side of the
instrument. If the temperature is higher than 20°C, cool the
Vessel as follows:
a. Fill the Vessel with cold water.
b. When the Temperature Controller reads 20°C, run
the Flush Procedure to drain the water.
10. Attach each Cubetainer hose to the Cubetainer and to its
specific port. Port A is used for Crude Fiber Acid solution.
Port B is used for Crude Fiber Base solution.
11. Open the Vessel Lid and insert the Bag Suspender with bags
into the Vessel and place the Bag Suspender Weight on top
of the empty 9th tray to keep the Bag Suspender submerged.
12. Follow the instructions on the ANKOM2000 display:
a. Select Crude Fiber.
b. Close Vessel Lid.
c. Confirm hot water is on (>50°C).
d. Press START.
13. When the Crude Fiber extraction and rinsing processes are
complete, open the Vessel Lid and remove the samples.
Gently press out excess water from the bags. Place the bags
in a 250ml beaker and add enough acetone to cover the bags
and soak for 3-5 minutes.
CAUTION4: Acetone is extremely flammable. Avoid static
electricity. A fume hood should be used at all times when
using acetone.
14. Remove the filter bags from the acetone and place them on a
wire screen to air-dry. Completely dry in an oven at 102 ±
2°C. (In most ovens the filter bags will be completely dry
within 2-4 hours.) NOTE: Do not place bags in the oven
until the acetone has completely evaporated.
15. Remove the filter bags from the oven and immediately place
them directly into a collapsible desiccant pouch and flatten
to remove any air. Cool to ambient temperature and weigh
the filter bags. NOTE: Do not use a conventional desiccator
container for this step.
16. Ash the entire filter bag/sample in a pre-weighed crucible
for 2 hours at 600 ± 15°C, cool in a conventional desiccator
and weigh to calculate loss of weight of organic matter (W3).
Crude Fiber Method
Method 1
11/26/13 pg. 3 of 3
Collaborative
Laboratory No. Rep
Whole
Corn
Cattle
Feed Alfalfa
Whole
Soy
Poultry
Starter
Calf
Starter
Swine
Feed
Horse
Feed
Soy
Meal
Pig
Starter
Dog
Food
% Crude Fiber
1 1 2.1 14.5 22.6 9.8 4.7 11.0 17.5 6.4 3.7 2.8 1.3
2 1.8 14.2 22.4 9.9 4.9 10.7 17.2 6.5 4.0 2.9 1.3
2 1 1.7 14.8 C 22.5 7.2 C 4.4 10.4 17.4 5.8 3.4 2.6 7.1 C
2 2.0 20.2 C 23.0 10.1 C 4.7 11.1 17.4 6.0 3.5 2.8 1.0 C
3 1 1.6 14.1 22.5 10.1 4.6 10.8 17.6 6.6 3.9 3.1 2.0
2 1.9 14.6 22.5 10.3 4.7 10.9 17.6 6.8 4.0 3.2 1.6
4 1 1.6 14.2 22.2 9.5 4.4 10.6 17.1 6.2 3.4 3.0 1.3
2 1.7 14.7 22.2 9.9 4.7 10.5 16.9 6.4 3.7 2.9 1.3
5 1 1.5 13.9 22.7 9.5 4.8 10.5 17.3 5.9 3.6 2.8 1.3
2 1.8 14.5 22.4 10.1 4.7 10.5 17.6 6.0 3.5 2.7 1.4
6 1 1.8 14.1 22.6 9.3 4.7 10.9 17.2 6.3 3.7 2.8 1.2
2 2.0 14.3 21.9 9.4 4.5 10.4 17.2 6.1 3.8 3.0 1.3
7 1 1.7 14.5 24.0 10.0 4.8 10.7 17.4 6.1 3.7 3.0 1.2
2 1.5 14.8 23.6 10.0 4.3 10.4 17.4 6.2 4.0 2.9 1.1
8 1 1.6 15.0 22.3 9.3 4.6 10.7 17.4 C 6.0 C 3.7 2.5 0.5
2 1.6 14.4 22.9 10.0 4.3 10.8 2.4 C 5.2 C 3.4 2.6 1.1
9 1 1.4 14.4 21.9 8.9 4.6 10.4 17.0 5.9 3.4 2.7 1.3
2 1.8 14.3 22.6 9.6 4.2 10.4 16.6 5.9 3.7 2.7 1.2
10 1 1.7 14.1 21.4 9.3 4.5 10.8 17.0 6.3 3.8 2.9 1.4
2 1.7 14.2 22.1 9.8 4.8 10.9 17.3 6.3 3.6 2.8 1.4
11 1 1.4 14.3 23.3 8.5 4.7 10.9 17.7 C 6.1 3.6 2.8 1.3
2 1.5 15.9 24.1 8.9 5.5 11.9 19.1 C 6.2 4.2 2.9 0.6
Mean 1.69 14.44 22.62 9.60 4.65 10.73 17.27 6.21 3.70 2.83 1.25
Official Method Laboratoriesa
% Crude Fiber
Central Analytical 1.8 14.5 23.0 10.2 4.4 9.3 G 14.7 G 6.8 2.9 1.9 G 3.4 G
Hahn Laboratories, Inc. 2.0 14.0 21.2 8.4 4.2 10.6 17.4 5.7 4.2 2.9 1.6
SDSU Olson Bio. Lab 2.4 14.2 23.8 10.1 4.6 10.8 17.4 6.8 4.1 2.8 1.3
Mean 2.05 14.23 22.67 9.57 4.40 10.70 17.40 6.43 3.73 2.85 1.45
Outliers: C-Chochran, G-Grubbs, DG-Double Grubbsa AOCS Official Method Ba 6-84, AOAC 962.09
Table 1. Results of the international collaborative study of the Filter Bag Technique for crude fiber compared
with three laboratories using an Official Crude Fiber Method.
Sample type
Whole
Corn
Cattle
Feed Alfalfa
Whole
Soy
Poultry
Starter
Calf
Starter
Swine
Feed
Horse
Feed
Soy
Meal
Pig
Starter
Dog
Food
Number of laboratories 11 10 11 10 11 11 9 10 11 11 10
Number of replicates 22 20 22 20 22 22 18 20 22 22 20
Overall FBT mean 1.69 14.44 22.62 9.60 4.65 10.73 17.27 6.21 3.70 2.83 1.25
Official Method meana
2.05 14.23 22.67 9.57 4.40 10.70 17.40 6.43 3.73 2.85 1.45
Sr 0.16 0.44 0.36 0.32 0.26 0.28 0.18 0.10 0.20 0.09 0.23
SR 0.19 0.44 0.67 0.48 0.27 0.33 0.28 0.27 0.22 0.17 0.31
RSDr, % 9.6 3.1 1.6 3.3 5.5 2.6 1.1 1.6 5.3 3.3 18.1
RSDR,% 11.4 3.1 2.9 5.0 5.8 3.1 1.6 4.3 6.0 6.0 24.5
r 0.46 1.23 1.00 0.88 0.72 0.80 0.51 0.27 0.55 0.26 0.64
R 0.54 1.23 1.86 1.34 0.75 0.94 0.78 0.75 0.62 0.48 0.86
HORRAT VALUE 3.07 1.14 1.18 1.75 1.82 1.11 0.62 1.42 1.83 1.75 6.34a Official Method AOCS Ba 6-84/AOAC 962.09
Table 2. Summary of the statistical analysis of the Filter Bag Technique crude fiber collaborative study, including
comparison with the Official Method.
Method 8 – determining Acid Detergent Lignin in beakers
3/13/13 pg. 1 of 2
Reagents
Sulfuric acid (72% by weight) – ANKOM Technology FSA72 or dilute reagent grade H2SO4 to a specific
gravity of 1634 g/L at 20°C (24.00N) by adding 1200 g H2SO4 to 350 ml H2O in a 1 L MCA volumetric flask
with cooling. Standardize this solution to 1634 g/L at 20°C specific gravity by removing solution and adding
H2O or H2SO4 as required.
Safety Precautions (see attached MSDS)
WARNINGS:
Acetone is highly flammable. Use fume hood when handling acetone and
avoid inhaling or contact with skin. Ensure that all the acetone has
evaporated before placing in the oven.
Wear rubber gloves and face shield when handling sulfuric acid. Always
add sulfuric acid to water. If acid contacts skin, wash with copious
amounts of water.
Apparatus
a) Filtration device – ANKOM Technology – F57 Filter Bags
b) Impulse bag sealer – ANKOM Technology – 1915 Heat Sealer
c) Desiccator – ANKOM Technology – Desiccant/MoistureStop pouch – X45
d) 2L & 3L Beaker
Procedure 1) Grind the sample to pass through a 1 mm screen (2mm screen when using a cyclone mill).
2) Weigh each Filter Bag (W1), record the weight, and tare the balance.
3) Add 0.5 g (±0.05 g) of air-dried sample (W2) directly into each Filter Bag.
4) Weigh and seal one (1) blank bag and include it in the digestion to determine the blank bag correction
(C1).
5) Seal the bags closed 4 mm from the open edge using the heat sealer.
6) Spread the sample uniformly inside each filter bag by flicking the bag to eliminate clumping.
7) Perform ADF determinations using Fiber Analyzer (See ADF Procedure).
8) After performing ADF determinations, place dried bags with samples into a 3 L beaker and completely
cover the bags with 72% H2SO4 (approximately 250 ml).
Bags must be completely dry and at ambient temperature before adding
concentrate acid. If moisture (even ambient moisture) is present in the
bags, heat generated by the H2SO4 and H20 reaction will adversely affect
the results.
9) Place a 2 L beaker inside the 3 L beaker to keep bags submerged. Agitate the bags at the start and at 30-
minute intervals by gently pushing and lifting the 2 L beaker up and down approximately 30 times.
10) After 3 hours pour off the H2SO4 and rinse with tap water to remove all acid.
If acid remains in the bags when they go into the oven, the samples will
burn, resulting in values that are higher than they should be.
11) Repeat rinses until pH paper shows neutral color when touching the bags. Rinse with approximately 250
ml of acetone for 3 minutes to remove the water. Handle the bags gently during rinsing. Fine lignin
particles can exit the filter if not handled carefully.
Method 8 – determining Acid Detergent Lignin in beakers
3/13/13 pg. 2 of 2
12) Dry the bags in an oven at 105 C for 2-4 hours.
WARNING – Do NOT place bags in an oven until all acetone has evaporated.
13) Remove the bags from the oven and place them directly into Desiccant/MoistureStop pouches and flatten
to remove air. Cool to ambient temperature and weigh the bags (W3).
14) Prepare each bag for the ash procedure. Extra Material
14.1) Fold each bag from bottom to top. Because the bags are wider at the
top than at the bottom, there will be a little extra material on each side
after the first fold.
14.2) Fold each bag from right to left. The extra material now lines up on
left side.
Extra Material
14.3) Heat seal the bag at the location of the extra material.
15) Ash the bags in pre-weighed crucibles (30 or 50 ml) at 525 C for 3 hours or until C-free. Cool and
calculate weight loss (W4).
16) Calculate blank bag ash correction (C2) using weight loss upon ignition of a blank bag sequentially run
through ADF and lignin steps.
17) Calculate percent ADL.
Where: W1 = Bag tare weight W2 = Sample weight W3 = Weight after extraction process W4 = Weight of Organic Matter (OM) (weight loss on ignition of bag and fiber residue) C1 = Blank bag correction (final oven-dried weight/original blank bag weight) C2 = Ash corrected blank bag (Loss of weight on ignition of bag/original blank bag)
DM = Dry Matter
ADL (as-received basis) = (W3 - (W1 x C1)) x 100
W2
ADLDM (DM basis) = (W3 - (W1 x C1)) x 100
W2 x DM
ADLOM (DM basis) = (W4 - (W1 x C2)) x 100
W2 x DM